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Chaperone proteins as single component reagents to assess antibody nonspecificity
Early stage assays that evaluate monoclonal antibody drug-like properties serve as valuable tools for selection of lead candidates. One liability for clinical development, off-target reactivity, is often assessed by binding to a mixture or panel of noncognate proteins. While robust, these mixes are...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627595/ https://www.ncbi.nlm.nih.gov/pubmed/28745541 http://dx.doi.org/10.1080/19420862.2017.1356529 |
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author | Kelly, Ryan L. Geoghegan, James C. Feldman, Jared Jain, Tushar Kauke, Monique Le, Doris Zhao, Jessie Wittrup, K. Dane |
author_facet | Kelly, Ryan L. Geoghegan, James C. Feldman, Jared Jain, Tushar Kauke, Monique Le, Doris Zhao, Jessie Wittrup, K. Dane |
author_sort | Kelly, Ryan L. |
collection | PubMed |
description | Early stage assays that evaluate monoclonal antibody drug-like properties serve as valuable tools for selection of lead candidates. One liability for clinical development, off-target reactivity, is often assessed by binding to a mixture or panel of noncognate proteins. While robust, these mixes are often ill-defined, and can suffer from issues such as lot-to-lot variability. In this study, we discovered in immunoprecipitation experiments that certain chaperones are present in one of these mixtures;we then explored the use of recombinant chaperone proteins as well-characterized agents to predict antibody nonspecificity. Antibody binding to the heat shock proteins HSP70, HSP90, or trigger factor all served as predictors of cross-interaction propensity, with HSP90 providing the greatest ability to predict antibody clearance rates in mouse. Individual chaperone binding correlates surprisingly closely with binding to complex cell extracts, with the exception of a few “false negatives” (assuming a complex cell extract as the “true” value). As defined reagents, these chaperone reagents present advantages for high throughput assays of nonspecificity. |
format | Online Article Text |
id | pubmed-5627595 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-56275952017-10-12 Chaperone proteins as single component reagents to assess antibody nonspecificity Kelly, Ryan L. Geoghegan, James C. Feldman, Jared Jain, Tushar Kauke, Monique Le, Doris Zhao, Jessie Wittrup, K. Dane MAbs Short Communication Early stage assays that evaluate monoclonal antibody drug-like properties serve as valuable tools for selection of lead candidates. One liability for clinical development, off-target reactivity, is often assessed by binding to a mixture or panel of noncognate proteins. While robust, these mixes are often ill-defined, and can suffer from issues such as lot-to-lot variability. In this study, we discovered in immunoprecipitation experiments that certain chaperones are present in one of these mixtures;we then explored the use of recombinant chaperone proteins as well-characterized agents to predict antibody nonspecificity. Antibody binding to the heat shock proteins HSP70, HSP90, or trigger factor all served as predictors of cross-interaction propensity, with HSP90 providing the greatest ability to predict antibody clearance rates in mouse. Individual chaperone binding correlates surprisingly closely with binding to complex cell extracts, with the exception of a few “false negatives” (assuming a complex cell extract as the “true” value). As defined reagents, these chaperone reagents present advantages for high throughput assays of nonspecificity. Taylor & Francis 2017-07-26 /pmc/articles/PMC5627595/ /pubmed/28745541 http://dx.doi.org/10.1080/19420862.2017.1356529 Text en © 2017 The Author(s). Published with license by Taylor & Francis Group, LLC http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way. |
spellingShingle | Short Communication Kelly, Ryan L. Geoghegan, James C. Feldman, Jared Jain, Tushar Kauke, Monique Le, Doris Zhao, Jessie Wittrup, K. Dane Chaperone proteins as single component reagents to assess antibody nonspecificity |
title | Chaperone proteins as single component reagents to assess antibody nonspecificity |
title_full | Chaperone proteins as single component reagents to assess antibody nonspecificity |
title_fullStr | Chaperone proteins as single component reagents to assess antibody nonspecificity |
title_full_unstemmed | Chaperone proteins as single component reagents to assess antibody nonspecificity |
title_short | Chaperone proteins as single component reagents to assess antibody nonspecificity |
title_sort | chaperone proteins as single component reagents to assess antibody nonspecificity |
topic | Short Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627595/ https://www.ncbi.nlm.nih.gov/pubmed/28745541 http://dx.doi.org/10.1080/19420862.2017.1356529 |
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